The objective of this project is to develop custom atomic force microscopy (AFM) systems capable of performing multi-scale corneal biomechanical characterization. Custom AFM instrumentation and techniques capable of corneal elasticity and viscoelasticity characterization at the nanoscale and macroscale length scales will be developed. This project will be applied to the field of corneal mechanics with a particular clinical focus to understand the mechanisms and effectiveness of corneal crosslinking, which is a promising treatment option for keratoconus. Using the nanoscale and bulk mechanical characterization results of crosslinked and normal corneas from the developed instrumentation and model, the effect of corneal crosslinking on corneal mechanical strength will be quantifies. The three specific aims of this project are: Aim 1: Develop instrumentation to enable the AFM characterization of the nanoscale and bulk corneal biomechanical properties Aim 2: Develop a model to determine corneal bulk properties from indentation depth analysis Aim 3: Quantify the effect of crosslinking on the biomechanical properties of the cornea The development of multi-scale corneal biomechanical characterization capability using atomic force microscopy will enhance knowledge regarding corneal biomechanics. Such insight will not only improve the ability to objectively measure the effectiveness of corneal crosslinking, but can also be expanded to other corneal diagnostic and treatment methods, glaucoma, and other ocular tissues.